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Finally, images were captured with fluorescent microscope

Alcohol would not fall under the category of an essential nutrient because not having it in your diet does not lead to any sort of deficiency

10.1242/dev.202449 [DOI] [PMC free article] [PubMed] [Google Scholar] Hamad M

Drug Des Dev Ther 13:3693

The plate was incubated in an Infinite F200 Pro plate reader (Tecan) with the lid on, maintaining a temperature of 37 C and shaking at 225 rpm for 24 h